17-(beta-hydroxyethylidene)-13-methyl-1, 2, 3, 6, 7, 8, 9, 10, 11, 12, 13, 14, 16, 17-tetradecahydro-15h-cyclopenta [a] phenanthren-3-one and esters thereof



United States Patent Office 2,704,768 Patented Mar. 22, 1955 17-(fi-HYDROXYETHYLIDENE) 13 NIETHYL- 1,2,3,6,7,8,9,10,11,12,13,14,16,17 TETRADECA- HYDRO 15H CYCLOPENTA [a] PHENAN- TI-IREN-3-ONE AND ESTERS THEREOF Frank B. Colton, Chicago, 11]., assignor to G. D. Searle & Co., Chicago, 11]., a corporation of Illinois No Drawing. Application May 25, 1953, Serial No. 357,377

5 Claims. (Cl. 260-397.47)

The present invention relates to a new group of organic polycyclic compounds and, more particularly to l7-(B- hydroxyethylidene) 13 methyl 1,2,3,6,7,8,9,l0,11,12,- 13,l4,16,17 tetradecahydro 15H cyclopentaEal-pheanthren-3-one and its esters. These compounds can be represented by the structural formula CHQOR (in CH: C I CH2 on. in than. C C C 0:

CH CH:

wherein R is a member of the class consisting of hydrogen, benzoyl and (lower alkyl)-CO radicals. Among the radicals which R can represent are acetyl, prop1onyl, iaifityryl, pentanoyl, hexanoyl, benzoyl, phenacetyl and the e. The claimed compositions are active horomonal agents. They are capable of maintaining life in adrenalectomized mammals. A special field of utility of these compounds consists in their use as starting material in the synthesis of adrenocortical hormones and especially of 2,17-dihydroxy 17 (6 hydroxyacetyl) 13 methyl 1,2,3,6,7,- 8,9,10,11,12,13,14,16,17 tetradecahydro 15H cyclopenta[a]phenanthren-3-one, a compound which combines valuable activities of the glucocorticoid adrenal hormones with an extremely potent life maintaining and growth promoting activity in adrenalectomizecl mammals. The compounds of this invention can be prepared by the method described in my copending application, Serial Number 286,611, filed May 7, 1952, issued as U. S. Patent 2,655,- 518, on October 13, 1953, of which the present application is a continuation-in-part. An alternative procedure is indicated in the experimental part below. The final step in the preparation of the claimed compounds can be represented by the following structural formulae.

CH: CH:

CHzBl' CHsOR be practiced without departing from the invention. In each of these examples temperatures are given in degrees MOB.

centigrade C.) and relative amounts of materials in parts by weight.

Example 1 A stirred solution of 10.6 parts of 3-methoxy-l3-methyl l,4,6,7,8,9,11,12,13,14,16,17 dodecahydro 15H- cyclopenta[alphenanthren-17-one in 700 parts of anhydrous ether and 45 parts of dry toluene is cooled to 0 C. and saturated with dry acetylene. While a slow stream of acetylene is passed through the reaction mixture, a solution of 20 parts of potassium t-amylate in parts of anhydrous pentanol is added in the course of 15 minutes with stirring. Passage of acetylene and stirring are continued for an additional 4V2 hours. After standing at 0 C. for 16 hours, the mixture is washed with aqueous ammonium chloride solution until the aqueous phase is neutral, then with water and saturated sodium chloride solution. The organic layer is dried over anhydrous sodium sulfate, filtered and concentrated under vacuum to a residue of about 250 parts. 500 parts of petroleum ether are added and, after standing at 0 C. for an hour, the mixture is filtered. The collected precipitate is recrystallized from ether. The resulting 3-methoxy-13- methyl 17 ethynyl l,4,6,7,8,9,11,12,13,14,16,17 dodecahydro 15H cyclopenta[a]phenanthren 17 ol melts at about l81182 C. The rotation as determined in a 1% chloroform solution is [a]D=-I65. An additional amount of this product can be obtained from the mother liquors by concentration under vacuum followed by addition of petroleum ether.

Example 2 To a refluxing solution of 47.5 parts of 3-methoxy-l3- methyl 17 ethynyl 1,4,6,7,8,9,l1,12,13,l4,16,l7 dodecahydro 15H cyclopenta [alphenanthren-l7-ol in 3200 parts of methanol and 1000 parts of water are added 240 parts of concentrated hydrochloric acid. Refluxing is continued for an additional 5 minutes after which the solution is maintained at room temperature for 15 minutes. Then 13,000 parts of water are added and the mixture is cooled to 0 C. After standing for several hours at that temperature, the mixture is filtered and the precipitate is dried and crystallized from ethyl acetate. The 13 methyl 17 ethynyl 17 hydroxy 1,2,3,6,7,- 8,9,l0,ll,l2,l3,14,16,17 tetradecahydro 15H cyclopenta[alphenanthren3-one thus obtained melts at about 202-204 C. The rotation, as determined in a 1% chloroform solution, is [aln=-22.5. The ultraviolet absorption spectrum of a methanolic solution shows a maximum at 241 millimicrons with a molecular extinction coeflicielrllt of 17,100. The compound has the structural form a CH III C Example 3 A solution of 53.7 parts of 13-methyl-17-ethynyl-l7-hydroxy 1,2,3,6,7,8,9,10,11,12,13,14,16,17 tetradecahydro 15H cyclopentaialphenanthren 3 one in 1500 parts of dioxane and 1000 parts of pyridine is reduced in an atmosphere of hydrogen over 30 parts of a catalyst containing 5% palladium on calcium carbonate. On absorption of one molecule of hydrogen the reduction is stopped and the mixture is filtered. The filtrate is concentrated under vacuum to about 500 parts, diluted with 3000 parts of ether and washed with normal hydrochloric acid until a Congo red test shows an acidic reaction. The solution is Washed successively with water, 5% sodium bicarbonate, water and saturated sodium chloride solution. The ether extract is dried over sodium sulfate, concentrated on the steam bath to about 500 parts and diluted with 800 arts of petroleum ether. After storage at 0 C. for 16 ours, the product is collected on a filter, dried and crystallized from a mixture of ethyl acetate and pe- Example 4 A solution of 47.3 parts of phosphorus tribromide in 645 parts of anhydrous ethanol-free chloroform is added dropwise to a solution of 142.9 parts of 13-methyl-17- vinyl 17 hydroxy 1,2,3,6,7,8,9,10,ll,12,13,14,l6,17- tetradecahydro l5H-cyclopenta[alphenanthren 3 one in 2250 parts of chloroform and parts of pyridine, maintained at -20 C. After standing at room temperature for 16 hours, the mixture is treated with chloroform and then successively with dilute hydrochloric acid, dilute sodium bicarbonate solution and finally with water. After drying over anhydrous sodium sulfate, the chloroform is stripped olT, leaving as a residue the 17-(p-bromoethylidene) 13 methyl l,2,3,6,7,8,9,10,l1,12,13,- 14,16,17 tetradecahydro 15H cyclopentaEalphenanthren-3-one. This compound has the structural formula CHsBt Example 5 45 parts of 17-(fi-bromoethylidene)-13-methyl-l,2,3,- 6,7,8,9,10,11,12,l3,14,16,17 tetradecahydro 15H cyclopenta[alphenanthren-B-one are treated with 400 parts of freshly fused potassium acetate and refluxed for 5 hours in 3200 parts of dry acetone. After cooling the precipitate is removed by filtration and the acetone is distilled in vacuum under nitrogen. The residue is extracted by refluxing with boiling petroleum ether and,

after stripping of the solvent in vacuo, the residue is chromatographed over 4500 parts of silica ,gel. .Elution with a 3% 'solution of ethyl acetate in benzene, eva'poration of' the solvent from the eluate and crystallization of the residue from aqueous acetone and petroleum ether yields 13 methyl 17 vinyl l,2,3,6,7,8,9,10,11,12,13,- 14 dodecahydro 15H cyclopenta[a]phenanthren '3- one, melting at about 100-101" C. The rotation of an 0.66% chloroform solution is z] =+l10.5'. The ultraviolet absorption spectrum of a methanolic solution shows a maximum at 237 millimicrons with a molecular extinction coclficient of 30,200. This compound has the structural formula Elution of the chromatography column with a 10% solution of ethyl acetate in benzene, evaporation of the solvent from the eluate and recrystallization of the residue from aqueous acetone yields the 17-(fl-acetoxyethylidene) 13 methyl 1,2,3,6,7,8,9,l0,11,12,l3,14,16,17- tetradecahydro 15H cyclopentalalphenanthren 3- one. This compound is obtained in two polymorphic crystalline forms, one melting at 49-50 C., the other melting at about 96-97 C. The rotation of a 1% chloroform solution is '[a] =+62.5. The ultraviolet absorption spectrum of a methanolic solution shows a maximum at 241 millimicrons with a molecular extinction coefiicient of 17,800. This compound has the structural formula CHaO-OC-CH:

Example 6 1000 parts of a 2-N potassium hydroxide solution in 75% aqueous methanol are treated with 45.9 parts of 17 (p acetoxyethylidene) l3 methyl 1,2,3,6,7,- 8,9,10,1l,l2,13,l4,l6,17 tetradecahydro 15H cyclopenta[a]phenanthren-3-one and refluxed for 17 minutes. Water is added to turbidity and the mixture is cooled to 0 C. The precipitate is collected on a filter, washed with water and dissolved in ethyl acetate. The solution is decolorized with charcoal, concentrated to one-third of its volume and treated with petroleum ether to induce crystallization. The l7-(fl-hydroxyethylidene)-13-methyl 1,2,3,6,7,8,9,10,11,12,13,14,16,17 tetradecahydro- 15H-cyclopenta[alphenanthren-3-one thus obtained melts at about 151-153 C. The identical product is obtained by alkaline hydrolysis in methanol of the 17-(B-bromoethylidene) 13 methyl 1,2,3,6,7,8,9,10,11,12,13,14,- 16,l7 tetradecahydro 15H cyclopenta[a]phenanthren-B-one obtained in Example 4. A mixed melting point test of the material obtained by these two methods shows no depression. A 1% chloroform solution shows an optical rotation [a] =+51. The product has the structural formula CHtOH Example 7 A mixture of 30 parts of l7-(p-bromoethylidene)-l3- methyl 1,2,3,6,7,8,9,10,11,12,13,14,16,17 tetradecahydro-l5H-cyclopenta[aJphenanthren-3-one and 40 parts of potassium benzoate is refluxed for 8 hours in 4000 parts of anhydrous acetone, cooled and filtered. The filtrate is freed from solvent by vacuum distillation under nitrogen. Chromatographic separation as in Example 5 on a silica gel column yields first the 13-methyl-l7- vinyl 1,2,3,6,7,8,9,10,11,12,13,14 dodecahydro 15H- cyclopenta[alphenanthren-3-one and then the 17-03- benzoyloxy) 13 methyl 1,2,3,6,7,8,9,10,11,12,13,- 14,16,17 tetradecahydro 15H cyclopenta[a]phenanthren-3-one. The ultraviolet absorption spectrum of the latter shows a maximum at 235 millimicrons with a molecular extinction coeflicient of 27,700. The infrared absorption spectrum shows maxima at 5.8, 6.0, 7.9 and Example 8 To a solution of parts of 17-(p-acetoxyethylidene)- 13 methyl 1,2,3,6,7,8,9,10,l1,12,13,14,l6,17 tetradecahydro 15H cyclopentalalphenanthren 3 one in 200 parts of tertiary butanol are added 0.27 part of osmium tetroxide in 16 parts of tertiary butanol, followed immediately by 60 parts of a 3.27-N hydrogen peroxide solution in tertiary butanol. In the course of the following two hours, a solution of 1.25 parts of osmium tetroxide in 80 parts of tertiary butanol is added. After standing at room temperature for 24 hours, the mixture is treated with 1500 parts of water and concentrated in vacuum at room temperature until about 320 parts of distillate have been collected. The residue is extracted with ethyl acetate and the extract is washed with water, dried over sodium sulfate, filtered, and evaporated to dryness. The residue is taken up in 1000 parts of methanol and refluxed for minutes with a solution of 9 parts of sodium sulfite in 200 parts of water. The reaction mixture is concentrated to about one-half of its original volume under nitrogen and extracted with ethyl acetate. This extract is washed with water, dried over sodium sulfate and evaporated. The residue contains a mixture of 13 methyl l7 glycolyl 17 hydroxy- 1,2,3,6,7,8,9,10,11,12,l3,14,l6,17 tetradecahydro 15H- cyclopentataJphenanthren 3 one and 17 (cafi dihydroxyethyl) 17 hydroxy 13 methyl 1,2,3,6,7,8,- 9,10,1l,12,l3,l4,l6,l7 tetradecahydro 15H cyclopentatalphenanthren-3-one. These compounds have the structural formulae CHsOH In addition the mixture contains a third compound which is apparently 4,5-dihydroxy-13-methyl-17-(5-hydroxyethylidene)perhydro 15H cyclopentalalphenanthren-S-one of the structural formula OHIO-00G HaOH The above residue is dissolved in parts of pyridine and 35 parts of acetic anhydride and kept at room temperature for 15 hours. Ice and, 2 hours later, water is added and the mixture is extracted with ethyl acetate. This extract is washed with dilute hydrochloric acid, sodium bicarbonate and water. After drying over sodium sulfate, the extract is evaporated under vacuum and the residue is chromatographed over 250 parts of silica gel. The column is eluted first with 1500 parts of a 10% solution of ethyl acetate in benzene. Elution with 500 parts each of a 10% and a 15% solution of ethyl acetate in benzene yields unreacted starting material. The column is next washed with an additional 500 parts of a 15% solution of ethyl acetate in benzene. Elution with a further 500-part portion of such a 15% solution and evaporation of the solvent yields a residue which, when crystallized from a mixture of ethyl acetate and petroleum ether and then from ether, forms crystals melting at about 187 C. This material gives a positive blue tetrazolium test and does not have a specific absorption maximum in the ultraviolet spectrum between 220 and 330 millimicrons. The infrared spectrum shows maxima at about 2.78, 5.78, 6.9, 7.3, 8.06, 8.79, 9.21, 9.5, 9.75, 10.3, 10.55, 10.8 and 11.3 microns. The compound is apparently 4 acetoxy 5 hydroxy l3 methyl 17- (B acetoxyethylidene)perhydro 15H cyclopentalalphenanthren-3-one of the structural formula CH2OOCCH:

CHsCOO OH CHzO-O C-CH! Further elution of the chromatography column with 2000 parts of a 20% solution and 1000 parts of a 30% solution of ethyl acetate in benzene and concentration of the eluate yields the l7-(a,B-diacetoxyethyl)-17-hydroxy- 13 methyl 1,2,3,6,7,8,9,10,1l,12,l3,14,16,17 tetradecahydro 15H cyclopentalalphenanthren 3 one which, recrystallized from ethyl acetate and petroleum ether, melts at about 194196 C. The rotation of an 0.28% chloroform solution is [a] =-|-13.5. This compound has the structural formula Example 9 1 part of 13methyl-17-(p-acetoxyacetyl)-17-hydroxy- 1,2,3,6,7,8,9,10,11,12,13,14,16,17-tetradecahydro 15H- cyclopenta[aJphenanthren-Ii-one is stirred with 5000 parts of citrated beef blood and 5000 parts of 0.85% aqueous sodium chloride solution. This solution is perfused three times through a surviving beef adrenal, cannulated through the vein and having a finely lacerated I surface. The perfusate is then extracted with isopropyl acetate. This extract is dried by azeotropic distillation and then concentrated to a residue of about 50 parts and diluted with 250 parts of benzene. On standing some dihydroxy 17 (B hydroxyacetyl)-l3-methyl- 1,2,3,6,7,8,9,l0,11,12,13,14,16,17-tetradecahydro 15H- cyclopenta [alphenanthren-3-one is obtained. Upon crystallization from a mixture of ethyl acetate and petroleum ether, using charcoal decolorization, it melts at 231-233 C. with decomposition. Additional yield is obtained by application of the mother liquors to a chromatography column containing 15 parts of silica gel. The column is washed with 50 parts of a solution of ethyl acetate in benzene, 100 parts of a 20% solution and 250 parts of a 50% solution of ethyl acetate in benzene. Subsequent elution with 450 parts of ethyl acetate, concentration of the eluate and recrystallization of the residue from a mixture of ethyl acetate and petroleum ether yields the principal quantity of the dihydroxy-l7-(fihydroxyacetyl)-13 methyl-1,2,3,6,7,8,9,l0,11,12,13,14,- 16,l7-tetradecahydro 15H-cyclopenta-[alphenanthren- 3-one, melting at 231-234 C. An 0.25% ethanol solution shows a rotation [a] =-I-130. The ultraviolet absorption hpectrum of a methanolic solution shows a maximum at 242 millimicrons with a molecular extinction coeflicient of 17,400. It gives a blue tetrazolium test and a red color with concentrated sulfuric acid.

I claim: 1. A compound of the structural formula berzizoyl and (lower alkyl)-CO- radicals.

CHaOH 3. A compound of the structural formula cmo-o C-(hwor alkyl) 01120-0 o-om cmo-oc-ctm References Cited in the file of this patent UNITED STATES PATENTS 2,528,897 Mayfield Nov. 7, 1950 

1. A COMPOUND OF THE STRUCTURAL FORMULA 